DNA sequencing using polymerase substrate-binding kinetics

Michael John Robert Previte, Chunhong Zhou, Matthew Kellinger, Rigo Pantoja, Cheng Yao Chen, Jin Shi, Beibei Wang, Amirali Kia, Sergey Etchin, John Vieceli, Ali Nikoomanzar, Erin Bomati, Christian Gloeckner, Mostafa Ronaghi, Molly Min He

Research output: Contribution to journalArticlepeer-review

7 Citations (Scopus)

Abstract

Next-generation sequencing (NGS) has transformed genomic research by decreasing the cost of sequencing. However, whole-genome sequencing is still costly and complex for diagnostics purposes. In the clinical space, targeted sequencing has the advantage of allowing researchers to focus on specific genes of interest. Routine clinical use of targeted NGS mandates inexpensive instruments, fast turnaround time and an integrated and robust workflow. Here we demonstrate a version of the Sequencing by Synthesis (SBS) chemistry that potentially can become a preferred targeted sequencing method in the clinical space. This sequencing chemistry uses natural nucleotides and is based on real-time recording of the differential polymerase/DNA-binding kinetics in the presence of correct or mismatch nucleotides. This ensemble SBS chemistry has been implemented on an existing Illumina sequencing platform with integrated cluster amplification. We discuss the advantages of this sequencing chemistry for targeted sequencing as well as its limitations for other applications.

Original languageEnglish
Article number5936
JournalNature communications
Volume6
DOIs
Publication statusPublished - 2015 Feb

All Science Journal Classification (ASJC) codes

  • Chemistry(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Physics and Astronomy(all)

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